The selected CD31 microbead-positive cells analyzed by flow cytometry with a panel of antibodies: [CD31 (PE. anti-mouse, eBioscience™), CD45 (FITC anti-mouse, eBioscience™) CD106 (PE, Rat-anti-mouse, BD-Pharmingen™), CD144 (APC, anti-mouse, eBioscience™), and endothelial selective adhesion molecule (ESAM) (APC, anti-mouse, Biolegend®) antibodies]. 500,000 cells/ml were collected per tube, washed with PBS, centrifuged at 400g for 5 min at room temperature (×2), incubated with the recommended dilution of antibodies, stored at 4°C for 1 h, and analyzed by flow cytometry (BD LSRFortessa™) within 24 h. The lower threshold uses to exclude debris and the live cells with gating (20,000 cells) according to forward scatter (FSC) × side scatter (SSC), followed by sections containing the antibodies. The data retrieve from the flow cytometry software and analyze by FlowJo software version 7.6.2.
Cd31 pe anti mouse
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
CD31 (PE, anti-mouse) is a laboratory reagent used for flow cytometric analysis. It is a fluorophore-conjugated antibody that binds to the CD31 antigen, also known as platelet endothelial cell adhesion molecule (PECAM-1), which is expressed on the surface of various cell types, including endothelial cells and some leukocytes. The PE (phycoerythrin) fluorescent label allows for the detection and quantification of CD31-positive cells.
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Lab products found in correlation
Lsrfortessa, by BD (2 mentions)
Pe anti mouse cd29, by Thermo Fisher Scientific (2 mentions)
Apc anti mouse ki 67, by BioLegend (1 mentions)
Pe anti mouse cd45, by BioLegend (1 mentions)
Anti mouse cd11b pe, by Thermo Fisher Scientific (1 mentions)
Pe anti mouse sca 1, by BioLegend (1 mentions)
Pe anti mouse cd34, by BioLegend (1 mentions)
Anti mouse cd44 pe, by Thermo Fisher Scientific (1 mentions)
Pe anti mouse cd3, by BioLegend (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
Trypsin, by Biosharp (1 mentions)
4 protocols using cd31 pe anti mouse
1
Flow Cytometry Analysis of Endothelial Cell Markers
2
Multiparametric Flow Cytometry Analysis of Endothelial Cells
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The selected CD31 positive cells analyzed by flow cytometry with a panel of antibodies; [CD31 (PE. antimouse, eBioscience™), CD45 (FITC anti-mouse, eBioscience™) CD106 (PE, Rat-anti-mouse, BD-Pharmingen™), CD144 (APC, anti-mouse, eBioscience™) and ESAM (APC, anti-mouse, Biolegend®) antibodies]. 500,000cells/ml were collected per tube, washed with PBS, centrifuge at 400 g for 5 minutes at room temperature (x2), incubated with the recommended dilution of antibodies, store at 4 0 C for one hour, and analyzed by flow cytometry (BD LSRFortessa ™) within 24 hours. The lower threshold uses to exclude debris and the live cells with gating (20,000 cells) according to FSC x SSC, followed by sections containing the antibodies. The data retrieve from the flow cytometry software and analyze by flow Jo software version 7.6.2.
3
Flow Cytometry for Cell Surface Markers
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For flow-cytometric analysis, cells were trypsinised, resuspended in FACS buffer (1% FBS in PBS+1 mM CaCl2+1 mM MgCl2) and labelled with one of the following antibodies (all used at 1:200 and, unless stated otherwise, purchased from eBioscience, Hatfield, UK): PE-anti-mouse Flk1, PE-anti-mouse CD49a (Cambridge Bioscience, Cambridge, UK); PE-anti-mouse CD49b; PE-anti-mouse CD49e; PE-anti-mouse CD51; PE-anti-mouse CD29; PE-anti-mouse CD61; PE-anti-mouse integrin beta 5; PE-anti-mouse CD31; FITC-anti-mouse ICAM2; PE-anti-mouse VECAD; appropriate PE/FITC-labelled isotype-matched controls were from eBioscience. In the case of Flk1 analysis, cells were stimulated with 30 ng/ml VEGF at 37°C over a 60-min time course before trypsinisation.
4
Flow Cytometry Analysis of MSC Surface Markers
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Characterization of the surface markers of cells was performed by using flow cytometry. MSCs digested in a dish with 0.25% Trypsin (Biosharp) were suspended in staining buffer (PBS, 1% FBS) and 1 × 105 cells/sample were incubated with fluorescently labeled antibodies (all 1:200) for 30 min at 4 °C. The antibodies used for flow cytometry were as follows: PE anti-mouse CD29 (eBioscience), PE anti-mouse Sca-1 (Biolegend), PE anti-mouse CD140a (eBioscience), PE anti-mouse CD44 (eBioscience), PE anti-mouse CD73 (eBioscience), PE anti-mouse MHC II (eBioscience), PE anti-mouse CD45 (Biolegend), PE anti-mouse CD31 (eBioscience), PE anti-mouse CD34 (Biolegend), PE anti-mouse CD11B (eBioscience), PE anti-mouse ICAM-1 (Biolegend), and BV421 anti-mouse VCAM-1 (BD Pharmingen). For intracellular staining, cells stained with cell surface antibodies (PE anti-mouse CD3 (Biolegend)) were fixed, permeabilized using foxp3/transcription factor concentrate and diluent kit set prior to incubation with antibodies directed at intracellular antigens (APC anti-mouse KI-67 (Biolegend)). Fluorescence intensity was measured by flow cytometry (Cytoflex, Beckman Coulter). Data were analyzed with FlowJo v10 software and Cytexpert software.
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